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Preface to the Fifth Edition. <p>Important and Useful Equations for HPLC.</p> <p><b>1 Introduction.</b></p> <p>1.1 HPLC: A powerful separation method.</p> <p>1.2 A first HPLC experiment.</p> <p>1.3 Liquid chromatographic separation modes.</p> <p>1.4 The HPLC instrument.</p> <p>1.5 Safety in the HPLC laboratory.</p> <p>1.6 Comparison between high-performance liquid chromatography and gas chromatography.</p> <p>1.7 Comparison between high-performance liquid chromatography and capillary electrophoresis.</p> <p>1.8 Units for pressure, length and viscosity.</p> <p>1.9 Scientific journals.</p> <p>1.10 Recommended books.</p> <p><b>2 Theoretical Principles.</b></p> <p>2.1 The chromatographic process.</p> <p>2.2 Band broadening.</p> <p>2.3 The chromatogram and its purport.</p> <p>2.4 Graphical representation of peak pairs with different degree of resolution.</p> <p>2.5 Factors affecting resolution.</p> <p>2.6 Extra-column volumes (dead volumes).</p> <p>2.7 Tailing.</p> <p>2.8 Peak capacity and statistical resolution probability.</p> <p>2.9 Effects of temperature in HPLC.</p> <p>2.10 The limits of HPLC.</p> <p>2.11 How to obtain peak capacity.</p> <p><b>3 Pumps.</b></p> <p>3.1 General requirements.</p> <p>3.2 The short-stroke piston pump.</p> <p>3.3 Maintenance and repair.</p> <p>3.4 Other pump designs.</p> <p><b>4 Preparation of Equipment up to Sample Injection.</b></p> <p>4.1 Selection of the mobile phase.</p> <p>4.2 Preparation of the mobile phase.</p> <p>4.3 Gradient systems.</p> <p>4.4 Capillary tubing.</p> <p>4.5 Fittings.</p> <p>4.6 Sample injectors.</p> <p>4.7 Sample solution and sample volume.</p> <p><b>5 Solvent Properties.</b></p> <p>5.1 Table of organic solvents.</p> <p>5.2 Solvent selectivity.</p> <p>5.3 Miscibility.</p> <p>5.4 Buffers.</p> <p>5.5 Shelf life of mobile phases.</p> <p>5.6 The mixing cross.</p> <p><b>6 Detectors.</b></p> <p>6.1 General.</p> <p>6.2 UV detectors.</p> <p>6.3 Refractive index detectors.</p> <p>6.4 Fluorescence detectors.</p> <p>6.5 Electrochemical (amperometric) detectors.</p> <p>6.6 Light-scattering detectors.</p> <p>6.7 Other detectors.</p> <p>6.8 Multiple detection.</p> <p>6.9 Indirect detection.</p> <p>6.10 Coupling with spectroscopy.</p> <p><b>7 Columns and Stationary Phases.</b></p> <p>7.1 Columns for HPLC.</p> <p>7.2 Precolumns.</p> <p>7.3 General properties of stationary phases.</p> <p>7.4 Silica.</p> <p>7.5 Chemically modified silica.</p> <p>7.6 Styrene-divinylbenzene.</p> <p>7.7 Some other stationary phases.</p> <p>7.8 Column care and regeneration.</p> <p><b>8 HPLC Column Tests.</b></p> <p>8.1 Simple tests for HPLC columns.</p> <p>8.2 Determination of particle size.</p> <p>8.3 Determination of breakthrough time.</p> <p>8.4 The test mixture.</p> <p>8.5 Dimensionless parameters for HPLC column characterization.</p> <p>8.6 The van Deemter equation from reduced parameters and its use in column diagnosis.</p> <p>8.7 van Deemter curves and other coherences.</p> <p>8.8 Diffusion coefficients.</p> <p><b>9 Adsorption Chromatography: Normal-Phase Chromatography.</b></p> <p>9.1 What is adsorption?.</p> <p>9.2 The eluotropic series.</p> <p>9.3 Selectivity properties of the mobile phase.</p> <p>9.4 Choice and optimization of the mobile phase.</p> <p>9.5 Applications.</p> <p><b>10 Reversed-Phase Chromatography.</b></p> <p>10.1 Principle.</p> <p>10.2 Mobile phases in reversed-phase chromatography.</p> <p>10.3 Solvent selectivity and strength.</p> <p>10.4 Stationary phases.</p> <p>10.5 Method development in reversed-phase chromatography.</p> <p>10.6 Applications.</p> <p>10.7 Hydrophobic interaction chromatography.</p> <p><b>11 Chromatography with Chemically Bonded Phases.</b></p> <p>11.1 Introduction.</p> <p>11.2 Properties of some stationary phases.</p> <p>11.3 Hydrophilic interaction chromatography.</p> <p><b>12 Ion-Exchange Chromatography.</b></p> <p>12.1 Introduction.</p> <p>12.2 Principle.</p> <p>12.3 Properties of ion exchangers.</p> <p>12.4 Influence of the mobile phase.</p> <p>12.5 Special possibilities of ion exchange.</p> <p>12.6 Practical hints.</p> <p>12.7 Applications.</p> <p><b>13 Ion-Pair Chromatography.</b></p> <p>13.1 Introduction.</p> <p>13.2 Ion-pair chromatography in practice.</p> <p>13.3 Applications.</p> <p>13.4 Appendix: UV detection using ion-pair reagents.</p> <p><b>14 Ion Chromatography.</b></p> <p>14.1 Principle.</p> <p>14.2 Suppression techniques.</p> <p>14.3 Phase systems.</p> <p>14.4 Applications.</p> <p><b>15 Size-Exclusion Chromatography.</b></p> <p>15.1 Principle.</p> <p>15.2 The calibration chromatogram.</p> <p>15.3 Molecular mass determination by means of size-exclusion chromatography.</p> <p>15.4 Coupled size-exclusion columns.</p> <p>15.5 Phase systems.</p> <p>15.6 Applications.</p> <p><b>16 Affinity Chromatography.</b></p> <p>16.1 Principle.</p> <p>16.2 Affinity chromatography as a special case of HPLC.</p> <p>16.3 Applications.</p> <p><b>17 Choice of Method.</b></p> <p>17.1 The various possibilities.</p> <p>17.2 Method transfer.</p> <p><b>18 Solving the Elution Problem.</b></p> <p>18.1 The elution problem.</p> <p>18.2 Solvent gradients.</p> <p>18.3 Column switching.</p> <p>18.4 Comprehensive two-dimensional HPLC.</p> <p>18.5 Optimization of an isocratic chromatogram using four solvents.</p> <p>18.6 Optimization of the other parameters.</p> <p>18.7 Mixed stationary phases.</p> <p><b>19 Analytical HPLC.</b></p> <p>19.1 Qualitative analysis.</p> <p>19.2 Trace analysis.</p> <p>19.3 Quantitative analysis.</p> <p>19.4 Recovery.</p> <p>19.5 Peak-height and peak-area determination for quantitative analysis.</p> <p>19.6 Integration errors.</p> <p>19.7 The detection wavelength.</p> <p>19.8 Derivatization.</p> <p>19.9 Unexpected peaks: Ghost and system peaks.</p> <p><b>20 Quality Assurance.</b></p> <p>20.1 Is it worth the effort?.</p> <p>20.2 Verification with a second method.</p> <p>20.3 Method validation.</p> <p>20.4 Standard operating procedures.</p> <p>20.5 Measurement uncertainty.</p> <p>20.6 Qualifications, instrument test and system suitability test.</p> <p>20.7 The quest for quality.</p> <p><b>21 Preparative HPLC.</b></p> <p>21.1 Problem.</p> <p>21.2 Preparative HPLC in practice.</p> <p>21.3 Overloading effects.</p> <p>21.4 Fraction collection.</p> <p>21.5 Recycling.</p> <p>21.6 Displacement chromatography.</p> <p><b>22 Separation of Enantiomers.</b></p> <p>22.1 Introduction.</p> <p>22.2 Chiral mobile phases.</p> <p>22.3 Chiral liquid stationary phases.</p> <p>22.4 Chiral solid stationary phases.</p> <p>22.5 Indirect separation of enantiomers.</p> <p><b>23 Special Possibilities.</b></p> <p>23.1 Micro, capillary and chip HPLC.</p> <p>23.2 High-speed and super-speed HPLC.</p> <p>23.3 Fast separations at 1000 bar: UHPLC.</p> <p>23.4 HPLC with supercritical mobile phases.</p> <p>23.5 HPLC with superheated water.</p> <p>23.6 Electrochromatography.</p> <p><b>24 Appendix 1: Applied HPLC Theory.</b></p> <p><b>25 Appendix 2: How to Perform the Instrument Test.</b></p> <p>25.1 Introduction.</p> <p>25.2 Test sequence.</p> <p>25.3 Preparations.</p> <p>25.4 Pump test.</p> <p>25.5 UV detector test.</p> <p>25.6 Autosampler test.</p> <p>25.7 Column oven test.</p> <p>25.8 Equations and calculations.</p> <p>25.9 Documentation.</p> <p><b>26 Appendix 3: Troubleshooting.</b></p> <p>26.1 Pressure problems.</p> <p>26.2 Leak in the pump system.</p> <p>26.3 Deviating retention times.</p> <p>26.4 Injection problems.</p> <p>26.5 Baseline problems.</p> <p>26.6 Peak shape problems.</p> <p>26.7 Problems with light-scattering detectors.</p> <p>26.8 Other causes.</p> <p>26.9 Instrument test.</p> <p><b>27 Appendix 4: Column Packing.</b></p> <p><b>Index of Separations.</b></p> <p><b>Subject Index.</b></p>

"This book is very much ‘‘an aid to this end'' and should be number one on every young chromatographer's book list. The publishers too have played their part in producing a book that is pleasing on the eye and a pleasure to read." (Chromatographia, 1 December 2010)<br />

<p><strong>Veronika R. Meyer</strong>, Department of Biocompatible Materials, Switzerland Swiss Federal Laboratories for Materials Testing and Research, Switzerland.

<b>Jump into the HPLC adventure!</b> <p>Three decades on from publication of the 1^st German edition of Veronika Meyer's book on HPLC, this classic text remains one of the few titles available on general HPLC aimed at practitioners.</p> <p>New sections on the following topics have been included in this fifth edition:</p> <ul type="disc"> <li>Comparison of HPLC with capillary electrophoresis</li> <li>How to obtain peak capacity</li> <li>van Deemter curves and other coherences</li> <li>Hydrophilic interaction chromatography</li> <li>Method transfer</li> <li>Comprehensive two-dimensional HPLC</li> <li>Fast separations at 1000 bar</li> <li>HPLC with superheated water</li> </ul> <p>In addition, two chapters on the instrument test and troubleshooting in the appendix have been updated and expanded by Bruno E. Lendi, and many details have been improved and numerous references added.</p> <p>A completely new chapter is presented on quality assurance covering:</p> <ul type="disc"> <li>Is it worth the effort?</li> <li>Verification with a second method</li> <li>Method validation</li> <li>Standard operating procedures</li> <li>Measurement uncertainty</li> <li>Qualifications, instrument test, and system suitability test</li> <li>The quest for quality</li> </ul> <p><b>Reviews of earlier editions</b></p> <p>"That this text is written by an expert in both the practice and teaching of HPLC is evident from the first paragraph....not only an enjoyable, fascinating and easy read, but a truly excellent text that has and will serve many teachers, students and practitioners very well." —<i>The Analyst</i></p> <p>“…provides essential information on HPLC for LC practitioners in academia, industry, government, and research laboratories…a valuable introduction." - <i>American Journal of Therapeutics</i></p>

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